Rho激酶抑制剂联合葛根素通过抑制TGF-β1/Smads信号通路和炎症反应来减轻缺氧诱导肺动脉高压对肺血管的损伤

Alleviation of pulmonary vasculature injury induced by hypoxia-induced pulmonary hypertension via Rho kinase inhibitor combined with puerarin by inhibiting TGF-β1/Smads signaling pathway and inflammatory response

目的探讨Rho激酶抑制剂法舒地尔联合葛根素对缺氧诱导的肺动脉高压(hypoxia-induced pulmonary hypertension,HPH)大鼠TGF-β1/Smads通路及其炎症因子的影响。方法将60只SD大鼠按照随机数字表法分为5组:对照组、模型组、葛根素组、法舒地尔组和法舒地尔与葛根素联合治疗组,常压缺氧舱内缺氧处理构建缺氧诱导的肺动脉高压大鼠模型。采用测压导管测定血流动力学指标RVSP、mPAP、PASP和PAWP,并计算右心室肥厚指数RVHI。采用HE染色法观察肺动脉血管平滑肌层的损伤情况。采用Western blot法检测TGF-β1/Smads通路相关因子的表达情况。采用qRT-PCR法检测炎症相关因子的表达情况。采用免疫组化法检测Gal-3蛋白的表达情况。结果与模型组、葛根素组和法舒地尔组相比较,联合治疗组肺动脉血管壁厚度和炎性浸润程度下降,RVSP、mPAP、PASP、PAWP和RVHI下降(P<0.05),TGF-β1/Smads通路相关蛋白、炎症相关因子和Gal-3蛋白的表达量下降(P<0.05)。结论Rho激酶抑制剂法舒地尔联合葛根素能很好的改善缺氧性肺动脉高压对肺血管的损伤,为缺氧性肺动脉高压的临床治疗提供理论依据。

Objective To investigate the effect of Rho kinase inhibitor fasudil combined with puerarin on the TGF-β1/Smads pathway and inflammatory factors in hypoxia-induced pulmonary hypertension(HPH).Methods Sixty SD rats were divided into Sham group,model group,Puerarin group,Fasudil group and Fasudil and Puerarin combined treatment group.HPH rat model was constructed by hypoxia treatment in an atmospheric hypoxia chamber.Manometric catheters were used to measure the hemodynamic indicators RVSP,mPAP,PASP and PAWP,and the right ventricular hypertrophy index(RVHI)was calculated.HE staining was used to observe the injury of pulmonary artery vascular smooth muscle layer.Western blotting was used to detect the expression of related factors in the TGF-β1/Smads pathway.qRT-PCR was used to detect the expression of inflammation-related factors.The expression of Gal-3 protein was detected by immunohistochemistry.Results Pulmonary artery wall thickness and inflammatory infiltration of the combined treatment group declined;RVSP,mPAP,PASP,PAWP and RVHI decreased(P<0.05);TGF-β1/Smads pathway related factors,inflammation-related factors and Gal-3 protein expression decreased(P<0.05)when compared with those of the model group,Puerarin group and Fasudil group.Conclusion Rho kinase inhibitor fasudil combined with puerarin can improve the damage of hypoxic pulmonary hypertension to pulmonary artery,and provide a theoretical basis for clinical treatment of hypoxic pulmonary hypertension.